The relevance of Tim-3 polymorphisms and F protein to the outcomes of HCV infection

  • J. P. Pei
  • L. F. Jiang
  • X. W. Ji
  • W. XiaoEmail author
  • X. Z. DengEmail author
  • Z. X. Zhou
  • D. Y. Zhu
  • W. L. Ding
  • J. H. Zhang
  • C. J. Wang
  • K. Jing
Original Article


Hepatitis C virus (HCV) is one of the major causes of liver inflammation. The aim of this study was to investigate the associations of T-cell immunoglobulin and mucin domain-3 (Tim-3) polymorphisms and the alternate reading frame protein (F protein) with the outcomes of HCV infection. Three single-nucleotide polymorphisms (SNPs; rs10053538, rs12186731, and rs13170556) of Tim-3 were genotyped in this study, which included 203 healthy controls, 558 hepatitis C anti-F-positive patients, and 163 hepatitis C anti-F-negative patients. The results revealed that the rs12186731 CT and rs13170556 TC and CC genotypes were significantly less frequent in the anti-F-positive patients [odds ratio (OR) = 0.54, 95 % confidence interval (CI) = 0.35–0.83, p = 0.005; OR = 0.26, 95 % CI = 0.18–0.39, p < 0.001; and OR = 0.19, 95 % CI = 0.10–0.35, p < 0.001, respectively), and the rs13170556 TC genotype was more frequent in the chronic HCV (CHC) patients (OR = 1.70, 95 % CI = 1.20–2.40, p = 0.002). The combined analysis of the rs12186731 CT and rs13170556 TC/CC genotypes revealed a locus-dosage protective effect in the anti-F-positive patients (OR = 0.22, 95 % CI = 0.14–0.33, p trend < 0.001). Stratified analyses revealed that the frequencies of the rs12186731 (CT + TT) genotypes were significantly lower in the older (OR = 0.31, 95 % CI = 0.15–0.65, p = 0.002) and female (OR = 0.30, 95 % CI = 0.17–0.52, p < 0.001) subgroups, and rs13170556 (TC + CC) genotypes exhibited the same effect in all subgroups (all p < 0.001) in the anti-F antibody generations. Moreover, the rs13170556 (TC + CC) genotypes were significantly more frequent in the younger (OR = 1.86, 95 % CI = 1.18–2.94, p = 0.007) and female (OR = 2.38, 95 % CI = 1.48–3.83, p < 0.001) subgroups of CHC patients. These findings suggest that the rs12186731 CT and rs13170556 TC/CC genotypes of Tim-3 provide potential protective effects with the F protein in the outcomes of HCV infection and that these effects are related to sex and age.


HBsAg Seroclearance Alternate Reading Frame Protein 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Compliance with ethical standards


This study was funded by grants from the National Natural Science Foundation of China (grant nos. 81573213 and 81172724), the Natural Science Foundation of Jiangsu Province, China (grant nos. BK20151089 and BL2013021), and the Tian Qing Liver Disease Research Fund of the Chinese Hepatitis Foundation (grant no. CFHPC20132071).

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical approval

All procedures performed in the studies involving human participants were conducted in accordance with the ethical standards of the institutional and/or national research committee and in accordance with the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards.

Informed content

Informed consent was obtained from all individual participants included in this study.


National Natural Science Foundation of China (CN) (81573213); Xiaozhao Deng.

National Natural Science Foundation of China (CN) (81172724); Xiaozhao Deng.

Natural Science Foundation of Jiangsu Province, China (BK20151089); Xiaozhao Deng.

Natural Science Foundation of Jiangsu Province, China (BL2013021); not applicable.

Tian Qing Liver Disease Research Fund of Chinese Hepatitis Foundation (CFHPC20132071); Longfeng Jiang.


  1. 1.
    Shepard CW, Finelli L, Alter MJ (2005) Global epidemiology of hepatitis C virus infection. Lancet Infect Dis 5:558–567CrossRefPubMedGoogle Scholar
  2. 2.
    Hoofnagle JH (2002) Course and outcome of hepatitis C. Hepatology 36:S21–S29CrossRefPubMedGoogle Scholar
  3. 3.
    Moradpour D, Penin F, Rice CM (2007) Replication of hepatitis C virus. Nat Rev Microbiol 5:453–463CrossRefPubMedGoogle Scholar
  4. 4.
    Chisari FV (2005) Unscrambling hepatitis C virus–host interactions. Nature 436:930–932CrossRefPubMedGoogle Scholar
  5. 5.
    Frazier AD, Zhang CL, Ni L et al (2010) Programmed death-1 affects suppressor of cytokine signaling-1 expression in T cells during hepatitis C infection. Viral Immunol 23:487–495CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Nakamoto N, Cho H, Shaked A et al (2009) Synergistic reversal of intrahepatic HCV-specific CD8 T cell exhaustion by combined PD-1/CTLA-4 blockade. PLoS Pathog 5:e1000313CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Golden-Mason L, Palmer BE, Kassam N et al (2009) Negative immune regulator Tim-3 is overexpressed on T cells in hepatitis C virus infection and its blockade rescues dysfunctional CD4+ and CD8+ T cells. J Virol 83:9122–9130CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Monney L, Sabatos CA, Gaglia JL et al (2002) Th1-specific cell surface protein Tim-3 regulates macrophage activation and severity of an autoimmune disease. Nature 415:536–541CrossRefPubMedGoogle Scholar
  9. 9.
    Yan J, Zhang Y, Zhang JP et al (2013) Tim-3 expression defines regulatory T cells in human tumors. PLoS One 8:e58006CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Sabatos CA, Chakravarti S, Cha E et al (2003) Interaction of Tim-3 and Tim-3 ligand regulates T helper type 1 responses and induction of peripheral tolerance. Nat Immunol 4:1102–1110CrossRefPubMedGoogle Scholar
  11. 11.
    Wu W, Shi Y, Li SP et al (2012) Blockade of Tim-3 signaling restores the virus-specific CD8+ T-cell response in patients with chronic hepatitis B. Eur J Immunol 42:1180–1191CrossRefPubMedGoogle Scholar
  12. 12.
    Jost S, Moreno-Nieves UY, Garcia-Beltran WF et al (2013) Dysregulated Tim-3 expression on natural killer cells is associated with increased Galectin-9 levels in HIV-1 infection. Retrovirology 10:74CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Flecken T, Sarobe P (2015) Tim-3 expression in tumour-associated macrophages: a new player in HCC progression. Gut 64:1502–1503CrossRefPubMedGoogle Scholar
  14. 14.
    Cheng YQ, Ren JP, Zhao J et al (2015) Microrna-155 regulates interferon-gamma production in natural killer cells via Tim-3 signalling in chronic hepatitis C virus infection. Immunology 145:485–497CrossRefPubMedGoogle Scholar
  15. 15.
    Golden-Mason L, Waasdorp Hurtado CE, Cheng LL et al (2015) Hepatitis C viral infection is associated with activated cytolytic natural killer cells expressing high levels of T cell immunoglobulin- and mucin-domain-containing molecule-3. Clin Immunol 158:114–125CrossRefPubMedGoogle Scholar
  16. 16.
    Varaklioti A, Vassilaki N, Georgopoulou U et al (2002) Alternate translation occurs within the core coding region of the hepatitis C viral genome. J Biol Chem 277:17713–17721CrossRefPubMedGoogle Scholar
  17. 17.
    Samrat SK, Li W, Singh S et al (2014) Alternate reading frame protein (F protein) of hepatitis C virus: paradoxical effects of activation and apoptosis on human dendritic cells lead to stimulation of T cells. PLoS One 9:e86567CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Zhang L, Hao CQ, Miao L et al (2014) Role of Th1/Th2 cytokines in serum on the pathogenesis of chronic hepatitis C and the outcome of interferon therapy. Genet Mol Res 13:9747–9755CrossRefPubMedGoogle Scholar
  19. 19.
    Yue M, Deng X, Zhai X et al (2013) Th1 and Th2 cytokine profiles induced by hepatitis C virus F protein in peripheral blood mononuclear cells from chronic hepatitis C patients. Immunol Lett 152:89–95CrossRefPubMedGoogle Scholar
  20. 20.
    Dalagiorgou G, Vassilaki N, Foka P et al (2011) High levels of HCV core+ 1 antibodies in HCV patients with hepatocellular carcinoma. J Gen Virol 92:1343–1351CrossRefPubMedGoogle Scholar
  21. 21.
    Xiao W, Zhang Q, Deng XZ et al (2015) HCV F protein amplifies the predictions of IL-28B and CTLA-4 polymorphisms about the susceptibility and outcomes of HCV infection in Southeast China. Infect Genet Evol 34:52–60CrossRefPubMedGoogle Scholar
  22. 22.
    Han KH, Yoon KT (2008) New diagnostic method for liver fibrosis and cirrhosis. Intervirology 51:S11–S16CrossRefGoogle Scholar
  23. 23.
    Simmonds P, McOmish F, Yap PL et al (1993) Sequence variability in the 5′ non-coding region of hepatitis C virus: identification of a new virus type and restrictions on sequence diversity. J Gen Virol 74:661–668CrossRefPubMedGoogle Scholar
  24. 24.
    Baghbani-arani F, Roohvandv F, Aghasadeghi MR et al (2012) Expression and characterization of Escherichia coli derived hepatitis C virus ARFP/F protein. Mol Biol 46:226–235CrossRefGoogle Scholar
  25. 25.
    Kong J, Deng XZ, Wang ZC et al (2009) Hepatitis C virus F protein: a double-edged sword in the potential contribution of chronic inflammation to carcinogenesis. Mol Med Rep 2:461–469PubMedGoogle Scholar
  26. 26.
    Song HH, Ma SL, Cha ZS et al (2013) T-cell immunoglobulin and mucin-domain-containing molecule 3 genetic variants and HIV+ non-Hodgkin lymphomas. Inflammation 36:793–799Google Scholar
  27. 27.
    Li Z, Liu Z, Zhang G et al (2012) TIM3 gene polymorphisms in patients with chronic hepatitis B virus infection: impact on disease susceptibility and hepatocellular carcinoma traits. Tissue Antigens 80:151–157CrossRefPubMedGoogle Scholar
  28. 28.
    Li SF, Ren YJ, Peng DY et al (2015) Tim-3 genetic variations affect susceptibility to osteoarthritis by interfering with interferon gamma in CD4+ T cells. Inflammation 38:1857–1863CrossRefPubMedGoogle Scholar
  29. 29.
    Troesch M, Jalbert E, Canobio S et al (2005) Characterization of humoral and cell-mediated immune responses directed against hepatitis C virus F protein in subjects co-infected with hepatitis C virus and HIV-1. AIDS 19:775–784CrossRefPubMedGoogle Scholar
  30. 30.
    Bain C, Parroche P, Lavergne JP et al (2004) Memory T-cell-mediated immune responses specific to an alternative core protein in hepatitis C virus infection. J Virol 78:10460–10469CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Wu WB, Shao SW, Zhao LJ et al (2007) Hepatitis C virus F protein up-regulates c-myc and down-regulates p53 in human hepatoma HepG2 cells. Intervirology 50:341–346CrossRefPubMedGoogle Scholar
  32. 32.
    Cohen M, Bachmatov L, Ben-Ari Z et al (2007) Development of specific antibodies to an ARF protein in treated patients with chronic HCV infection. Dig Dis Sci 52:2427–2432CrossRefPubMedGoogle Scholar
  33. 33.
    Ndhlovu LC, Lopez-Vergès S, Barbour JD et al (2012) Tim-3 marks human natural killer cell maturation and suppresses cell-mediated cytotoxicity. Blood 119:3734–3743CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Kared H, Fabre T, Bédard N et al (2013) Galectin-9 and IL-21 mediate cross-regulation between Th17 and treg cells during acute hepatitis C. PLoS Pathog 9:e1003422. doi: 10.1371/journal.ppat.1003422 CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Mengshol JA, Golden-Mason L, Arikawa T et al (2010) A crucial role for Kupffer cell-derived galectin-9 in regulation of T cell immunity in hepatitis C infection. PLoS One 5:e9504. doi: 10.1371/journal.pone.0009504 CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Merani S, Chen WN, Elahi S (2015) The bitter side of sweet: the role of Galectin-9 in immunopathogenesis of viral infections. Rev Med Virol 25:175–186CrossRefPubMedGoogle Scholar
  37. 37.
    Cao BW, Zhu LZ, Zhu ST et al (2010) Genetic variations and haplotypes in TIM-3 gene and the risk of gastric cancer. Cancer Immunol Immunother 59:1851–1857CrossRefPubMedGoogle Scholar
  38. 38.
    Liao JY, Zhang Q, Liao Y et al (2014) Association of T-cell immunoglobulin and mucin domain-containing molecule 3 (Tim-3) polymorphisms with susceptibility and disease progression of HBV infection. PLoS One 9:e98280CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Emery J, Pick N, Mills EJ et al (2010) Gender differences in clinical, immunological, and virological outcomes in highly active antiretroviral-treated HIV-HCV coinfected patients. Patient Prefer Adherence 4:97–103PubMedPubMedCentralGoogle Scholar
  40. 40.
    Corsi DJ, Karges W, Thavorn K et al (2016) Influence of female sex on hepatitis C virus infection progression and treatment outcomes. Eur J Gastroenterol Hepatol 28:405–411. doi: 10.1097/MEG.0000000000000567 PubMedGoogle Scholar
  41. 41.
    Zhang J, Daley D, Akhabir L et al (2009) Lack of association of TIM3 polymorphisms and allergic phenotypes. BMC Med Genet 10:62CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • J. P. Pei
    • 1
  • L. F. Jiang
    • 2
  • X. W. Ji
    • 1
  • W. Xiao
    • 3
    Email author
  • X. Z. Deng
    • 1
    • 4
    Email author
  • Z. X. Zhou
    • 5
  • D. Y. Zhu
    • 6
  • W. L. Ding
    • 7
  • J. H. Zhang
    • 4
  • C. J. Wang
    • 4
  • K. Jing
    • 8
  1. 1.Department of Biochemistry and Molecular Biology, School of Basic MedicineNanjing Medical UniversityNanjingChina
  2. 2.Department of Infectious DiseasesThe First Affiliated Hospital with Nanjing Medical UniversityNanjingChina
  3. 3.School of Life Science and TechnologyChina Pharmaceutical UniversityNanjingChina
  4. 4.Huadong Research Institute for Medicine and BiotechnicsNo. 293, Zhongshan East RoadNanjingChina
  5. 5.Department of Clinical LaboratoryNanjing Second HospitalNanjingChina
  6. 6.Department of Infectious Diseases at Luoyang Central Hospital Affiliated to Zhengzhou UniversityLuoyangChina
  7. 7.Department of Clinical LaboratoryYixing People’s HospitalYixingChina
  8. 8.Faculty of Chemical EngineeringHuaiyin Institute of TechnologyHuai’anChina

Personalised recommendations